1-Amino-triazolo(1,5-A)pyridine-substituted Urea Derivative and Uses Thereof

ABSTRACT

Provided herein is a compound, 1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea, pharmaceutical compositions thereof and crystal forms thereof. Also provided are particles (e.g., nanoparticles) comprising the compound, and pharmaceutical compositions thereof that are mucus penetrating. Also provided herein are methods and kits for using the compound, and pharmaceutical compositions thereof for treating and/or preventing diseases associated with abnormal or pathological angiogenesis and/or aberrant signaling of a growth factor (e.g., vascular endothelial growth factor (VEGF)), such as proliferative diseases (e.g., cancers, benign neoplasms, inflammatory diseases, autoimmune diseases) and ocular diseases (e.g., macular degeneration, glaucoma, diabetic retinopathy, retinoblastoma, edema, uveitis, dry eye, blepharitis, and post-surgical inflammation) in a subject in need thereof.

BACKGROUND

Growth factors play an important role in angiogenesis,lymphangiogenesis, and vasculogenesis, and they regulate angiogenesis ina variety of processes. Undesirable or pathological angiogenesis isassociated with diseases including diabetic retinopathy, psoriasis,cancer, rheumatoid arthritis, atheroma, Kaposi's sarcoma, andhemangioma. Angiogenic ocular conditions represent the leading cause ofirreversible vision loss in developed countries. In the United States,for example, retinopathy of prematurity, diabetic retinopathy, andage-related macular degeneration are the principal causes of blindnessin infants, working age adults, and the elderly, respectively.

Therefore, there is a need for new therapeutic compounds for thetreatment of diseases associated with the aberrant signaling of growthfactors and diseases associated with angiogenesis, such as cancer,macular degeneration, and diabetic retinopathy.

SUMMARY

Disclosed herein are urea derivatives such as1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,which is shown in Formula (I) below,

and pharmaceutical compositions thereof, and methods useful in treatingand/or preventing diseases associated with abnormal angiogenesis and/oraberrant signaling of a growth factor (e.g., vascular endothelial growthfactor (VEGF)). Diseases that may be treated and/or prevented by thedisclosed compounds, pharmaceutical compositions, kits, uses, andmethods include proliferative diseases (e.g., cancers, benign neoplasms,inflammatory diseases, autoimmune diseases) and ocular diseases (e.g.,macular degeneration, glaucoma, diabetic retinopathy, retinoblastoma,edema, uveitis, dry eye, blepharitis, and post-surgical inflammation).

In one aspect, disclosed herein is a compound of Formula (I):

and pharmaceutically acceptable salts, solvates, hydrates, polymorphs,co-crystals, tautomers, stereoisomers, and prodrugs thereof.

In another aspect, disclosed herein is a crystal form of

wherein the crystal form has an X-ray powder diffraction (XRPD) patternwith peaks at about 10.5 (e.g., 10.54), about 14.4 (e.g., 14.35), about19.3 (e.g., 19.26), about 21.7 (e.g., 21.67), and about 24.7 (e.g.,24.72) degrees 29.

Also disclosed herein are pharmaceutical compositions comprising1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)ureaor a compound of Formula (I), or the crystal form, and apharmaceutically acceptable excipient or carrier. In certainembodiments, the pharmaceutical compositions described herein include aneffective amount of the compound disclosed herein. The pharmaceuticalcomposition may be useful for treating proliferative diseases (e.g.,cancers, benign neoplasms, inflammatory diseases, autoimmune diseases)and/or ocular diseases (e.g., macular degeneration, glaucoma, diabeticretinopathy, retinoblastoma, edema, uveitis, dry eye, blepharitis, andpost-surgical inflammation) in a subject in need thereof. Thepharmaceutical composition may also be useful for inhibiting abnormalangiogenesis and/or aberrant signaling of a growth factor in a subjector cell.

In some embodiments, the compound of Formula (I),1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)ureamay be intended for delivery in a subject's tissues having mucus (e.g.,eye, respiratory tract, gastrointestinal tract, genito-urinary tract),which is a viscoelastic and adhesive substance that traps most foreignobjects (e.g., microorganisms, particles, dust). In one aspect thecompound of Formula (I),1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is formulated into mucus penetrating particles or mucus penetratingcrystals (collectively, MPPs) suitable for administration (e.g., topicalor inhalation) to tissues of the subject having mucus (e.g., eye,respiratory tract, gastrointestinal tract, genito-urinary tract).

In another aspect, disclosed herein are particles comprising1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea.In certain embodiments, the particles are mucus penetrating. Theparticles may include a coating surrounding a core. The core maycomprise primarily the compound of Formula (I), or the core may be apolymeric core with the compound encapsulated in the polymer. In certainembodiments, the particles are nanoparticles (e.g., particles having anaverage diameter of at least about 10 nm and less than about 1 μm). Theparticles may be useful in delivering the compound of Formula (I) to asubject. In certain embodiments, the particles are capable of deliveringthe compound in or through mucus of a subject.

Another aspects relate to pharmaceutical compositions comprising acompound of Formula (I),1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,and/or a plurality of particles. In certain embodiments, thepharmaceutical compositions are useful in delivering the compound ofFormula (I) to a subject.

In other aspects, disclosed herein are pharmaceutical compositionscomprising a plurality of particles comprising (i) a core comprising1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt thereof, and (ii) a coating of asurface altering agent surrounding the core, wherein the surfacealtering agent is present on the outer surface of the core at a densityof at least 0.01 surface altering agent per nm², and optionally, atleast one pharmaceutically acceptable excipient or carrier. In someembodiments, the surface altering agent is a triblock copolymer of thestructure (hydrophilic block)-(hydrophobic block)-(hydrophilic block).In some aspects, the triblock copolymer is a PLURONIC® or a poloxamer, apoly(vinyl alcohol) that is less than 95% hydrolyzed, or a polysorbate.

In certain embodiments, the compound, particle, or pharmaceuticalcomposition is formulated to be mucus penetrating.

Other aspects disclosed herein relate to methods of treating and/orpreventing a disease associated with abnormal angiogenesis in a subjectin need thereof.

Other aspects disclosed herein relate to methods of treating and/orpreventing a disease associated with aberrant signaling of a growthfactor signaling pathway in a subject in need thereof.

In other aspects, disclosed herein are methods of inhibitingangiogenesis in a subject in need thereof.

In other aspects, provided herein are methods of inhibiting aberrantsignaling of a growth factor signaling pathway in a subject or cell. Incertain embodiments, the growth factor is associated with angiogenesis.In certain embodiments, the growth factor is VEGF.

The methods disclosed herein include administering to the subject aneffective amount of a compound of Formula (I) or pharmaceuticalcomposition disclosed herein. In certain embodiments, the effectiveamount is a therapeutically effective amount. In certain embodiments,the effective amount is a prophylactically effective amount.

In yet other aspects, provided herein are the compound of Formula (I)and pharmaceutical compositions for use in the treatment and/orprevention of a disease associated with abnormal angiogenesis and/orassociated with aberrant signaling of a growth factor signaling pathwayin a subject in need thereof.

The present application refers to various issued patent, publishedpatent applications, journal articles, and other publications, all ofwhich are incorporated herein by reference.

The details of one or more embodiments are set forth herein. Otherfeatures, objects, and advantages will be apparent from the DetailedDescription, the Figures, the Examples, and the Claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the X-ray powder diffraction (XRPD) pattern of crystalform I.

FIG. 2 depicts a differential scanning calorimetry (DSC) thermogram ofcrystal form I of Compound I.

FIG. 3 depicts a thermal gravim analysis (TGA) thermogram of crystalform I of Compound I.

FIG. 4 depicts the XRPD pattern of crystal form I after milling.

DETAILED DESCRIPTION Definitions

The following definitions are more general terms used throughout thepresent application.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and loweranimals. Examples of pharmaceutically salts are salts of an amino groupformed with inorganic acids such as hydrochloric acid, hydrobromic acid,phosphoric acid, sulfuric acid, and perchloric acid or with organicacids such as acetic acid, oxalic acid, maleic acid, tartaric acid,citric acid, succinic acid, or malonic acid or by using other methodsknown in the art such as ion exchange. Other pharmaceutically acceptablesalts include adipate, alginate, ascorbate, aspartate, benzenesulfonate,benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate,citrate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate,gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄ alkyl)₄ ⁻ salts.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, lower alkyl sulfonate, and aryl sulfonate.

The term “prodrugs” refer to compounds, including derivatives of thecompounds described herein, which have cleavable groups which areremoved in vivo to leave a parent compound that is pharmaceutically.Such examples include, but are not limited to, choline ester derivativesand the like, N-alkylmorpholine esters and the like. Other derivativesof the compounds have activity in both their acid and acid derivativeforms, but in the acid sensitive form often offer advantages ofsolubility, tissue compatibility, or delayed release in the mammalianorganism. Prodrugs include acid derivatives well known to practitionersof the art, such as, for example, esters prepared by reaction of theparent acid with a suitable alcohol, or amides prepared by reaction ofthe parent acid compound with a substituted or unsubstituted amine, oracid anhydrides, or mixed anhydrides. Simple aliphatic or aromaticesters, amides, and anhydrides derived from acidic groups pendant on thecompounds are particular prodrugs. In some cases it is desirable toprepare double ester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈alkynyl, aryl, C₇-C₁₂ substituted aryl, and C₇-C₁₂ arylalkyl esters ofthe compounds described herein may be preferred.

The terms “administer,” “administering,” or “administration,” as usedherein, refers to implanting, absorbing, ingesting, injecting, inhaling,or otherwise introducing the compound of Formula (I), or apharmaceutical composition thereof, in or on a subject.

As used herein, the terms “treatment,” “treat,” and “treating” refer topreventing, reversing, alleviating, delaying the onset of, or inhibitingthe progress of a disease described herein. In some embodiments,treatment may be administered after one or more signs or symptoms of thedisease have developed or have been observed. In other embodiments,treatment may be administered in the absence of signs or symptoms of thedisease. For example, treatment may be administered to a susceptiblesubject prior to the onset of symptoms (e.g., in light of a history ofsymptoms and/or in light of exposure to a pathogen). Treatment may alsobe continued after symptoms have resolved, for example, to delay orprevent recurrence.

As used herein, the terms “condition,” “disease,” and “disorder” areused interchangeably.

An “effective amount” of a compound described herein refers to an amountsufficient to elicit the desired biological response, i.e., treating thecondition. As will be appreciated by those of ordinary skill in thisart, the effective amount of a compound of Formula (I) may varydepending on such factors as the desired biological endpoint, thepharmacokinetics of the compound, the condition being treated, the modeof administration, and the age and health of the subject. An effectiveamount encompasses therapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound described herein isan amount sufficient to provide a therapeutic benefit in the treatmentof a condition or to delay or minimize one or more symptoms associatedwith the condition. The term “therapeutically effective amount” canencompass an amount that improves overall therapy, reduces or avoidssymptoms, signs, or causes of the condition, and/or enhances thetherapeutic efficacy of another therapeutic agent. In certainembodiments, a “therapeutically effective amount” of a compound orcomposition is the amount needed to inhibit angiogenesis in a subject.

A “prophylactically effective amount” of a compound described herein isan amount sufficient to prevent a condition, or one or more symptomsassociated with the condition or prevent its recurrence. Aprophylactically effective amount of a compound means an amount of atherapeutic agent, alone or in combination with other agents, whichprovides a prophylactic benefit in the prevention of the condition. Theterm “prophylactically effective amount” can encompass an amount thatimproves overall prophylaxis or enhances the prophylactic efficacy ofanother prophylactic agent.

As used herein, the term “growth factor” refers to a naturally occurringsubstance (e.g., a protein or a steroid hormone) capable of stimulatingcellular growth, proliferation, and cellular differentiation. Growthfactors may act as signaling molecules between cells and/or promote celldifferentiation and maturation.

As used herein, the term “vascular endothelial growth factor” or “VEGF”refers to a signal protein produced by cells that stimulatevasculogenesis and angiogenesis. VEGFs are a sub-family of growthfactors, i.e., the platelet-derived growth factor family ofcysteine-knot growth factors. VEGFs are important signaling proteinsinvolved in both vasculogenesis and angiogenesis. VEGFs' normal functionis to create new blood vessels during embryonic development, new bloodvessels after injury, muscle following exercise, and new vessels(collateral circulation) to bypass blocked vessels. When VEGF isoverexpressed, it can contribute to a range of diseases, such asproliferative diseases (e.g., cancer) and vascular diseases in theretina of the eye and other parts of the body. VEGFs include a number ofproteins from two families that result from alternate splicing of mRNAfrom a single, 8-exon, VEGF gene. Examples of VEGFs include, but are notlimited to, VEGF-related proteins such as placental growth factor (PGF),VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, and VEGF-F. The term “VEGF” alsoencompasses VEGF receptors (VEGFRs), such as VEGFR-1, VEGFR-2 andVEGFR-3. A VEGFR may be membrane-bound (mbVEGFR) or soluble (sVEGFR).

A “proliferative disease” refers to a disease that occurs due toabnormal growth or extension by the multiplication of cells. Aproliferative disease may be associated with: 1) the pathologicalproliferation of normally quiescent cells; 2) the pathological migrationof cells from their normal location (e.g., metastasis of neoplasticcells); 3) the pathological expression of proteolytic enzymes such asthe matrix metalloproteinases (e.g., collagenases, gelatinases, andelastases); or 4) the pathological angiogenesis as in proliferativeretinopathy and tumor metastasis. Exemplary proliferative diseasesinclude cancers (i.e., “malignant neoplasms”), benign neoplasms,angiogenesis, inflammatory diseases, and autoimmune diseases.

As used herein, the term “angiogenesis” refers to the physiologicalprocess through which new blood vessels form from pre-existing vessels.Angiogenesis is distinct from vasculogenesis, which is the de novoformation of endothelial cells from mesoderm cell precursors. The firstvessels in a developing embryo form through vasculogenesis, after whichangiogenesis is responsible for most blood vessel growth during normalor abnormal development. Angiogenesis is a vital process in growth anddevelopment, as well as in wound healing and in the formation ofgranulation tissue. However, angiogenesis is also a fundamental step inthe transition of tumors from a benign state to a malignant one, leadingto the use of angiogenesis inhibitors in the treatment of cancer.Angiogenesis may be chemically stimulated by angiogenic proteins, suchas growth factors (e.g., VEGF).

The terms “neoplasm” and “tumor” are used herein interchangeably andrefer to an abnormal mass of tissue wherein the growth of the masssurpasses and is not coordinated with the growth of a normal tissue. Aneoplasm or tumor may be “benign” or “malignant,” depending on thefollowing characteristics: degree of cellular differentiation (includingmorphology and functionality), rate of growth, local invasion, andmetastasis. A “benign neoplasm” is generally well differentiated, hascharacteristically slower growth than a malignant neoplasm, and remainslocalized to the site of origin. In addition, a benign neoplasm does nothave the capacity to infiltrate, invade, or metastasize to distantsites. Exemplary benign neoplasms include, but are not limited to,lipoma, chondroma, adenomas, acrochordon, senile angiomas, seborrheickeratoses, lentigos, and sebaceous hyperplasias. In some cases, certain“benign” tumors may later give rise to malignant neoplasms, which mayresult from additional genetic changes in a subpopulation of the tumor'sneoplastic cells, and these tumors are referred to as “pre-malignantneoplasms.” An exemplary pre-malignant neoplasm is a teratoma. Incontrast, a “malignant neoplasm” is generally poorly differentiated(anaplasia) and has characteristically rapid growth accompanied byprogressive infiltration, invasion, and destruction of the surroundingtissue. Furthermore, a malignant neoplasm generally has the capacity tometastasize to distant sites. The term “metastasis,” “metastatic,” or“metastasize” refers to the spread or migration of cancerous cells froma primary or original tumor to another organ or tissue and is typicallyidentifiable by the presence of a “secondary tumor” or “secondary cellmass” of the tissue type of the primary or original tumor and not ofthat of the organ or tissue in which the secondary (metastatic) tumor islocated. For example, a prostate cancer that has migrated to bone issaid to be metastasized prostate cancer and includes cancerous prostatecancer cells growing in bone tissue.

As used herein, the term “cancer” refers to a malignant neoplasm.

As used herein, the term “inflammatory disease” or “inflammation” refersto a disease caused by, resulting from, or resulting in inflammation.The term “inflammatory disease” may also refer to a dysregulatedinflammatory reaction that causes an exaggerated response bymacrophages, granulocytes, and/or T-lymphocytes leading to abnormaltissue damage and/or cell death. An inflammatory disease can be eitheran acute or chronic inflammatory condition and can result frominfections or non-infectious causes. Examples of inflammatory diseasesinclude, without limitation, autoimmune disorders, systemic lupuserythematosus, psoriasis, cystic fibrosis, rheumatoid arthritis,inflammatory arthritis, Sjogren's syndrome, inflammatory bowel disease,Crohn's disease, ulcerative colitis, pneumonia, respiratory tractinflammation, adult respiratory distress syndrome (ARDS), asthma,allograft rejection, and vaginitis. Ocular inflammatory diseasesinclude, but are not limited to, allergy of the eye, uveitis (e.g.,anterior uveitis, intermediate uveitis, and posterior uveitis),conjunctivitis, panuveitis, cyclitis, scleritis, episcleritis, opticneuritis, retrobulbar optic neuritis, keratitis (e.g., immune keratitisand infectious keratitis), blepharitis, corneal ulcer, conjunctivalulcer and symptoms caused by them, ocular inflammatory diseases causedby ocular disorders, ocular inflammatory diseases caused by a physicalinjury, post-surgical inflammation, and dry eye (e.g., dry eyesyndrome).

As used herein, an “autoimmune disease” refers to a disease arising froman inappropriate immune response of the body of a subject againstsubstances and tissues normally present in the body. In other words, theimmune system mistakes some part of the body as a pathogen and attacksits own cells. This may be restricted to certain organs (e.g., inautoimmune thyroiditis) or involve a particular tissue in differentplaces (e.g., Goodpasture's disease which may affect the basementmembrane in both the lung and kidney).

The term “ocular disease” or “ocular disorder” refers to any eye diseaseand/or disorder. For example, ocular diseases can be disorders of theeyelid, lacrimal system and orbit, disorders of conjunctiva, disordersof sciera, cornea, iris and ciliary body, disorders of choroid andretina, glaucoma, disorders of optic nerve and visual pathways, occularyinflammatory diseases, or disorders of ocular muscles. Additionally,ocular disease can also refer to discomfort following injury, surgery,or laser treatment. Diseases and disorders of the eye include, but arenot limited to, macular degeneration, dry eye syndrome, uveitis,allergic conjunctivitis, glaucoma, and ocular rosacea. Dry eye syndrome(DES), otherwise known as keratoconjunctivitis sicca (KCS), keratitissicca, sicca syndrome, or xerophthalmia, is an eye disease caused bydecreased tear production or increased tear film evaporation commonlyfound in humans and some animals

The term “age-related macular degeneration” or “AMD” is an oculardisease which usually affects older adults and results in a loss ofvision in the center of the visual field (the macula) because of damageto the retina. It occurs in “dry” and “wet” forms. It is a major causeof blindness and visual impairment in older adults (>50 years). Maculardegeneration can make it difficult or impossible to read or recognizefaces, although enough peripheral vision remains to allow otheractivities of daily life. In the dry (nonexudative) form, cellulardebris called drusen accumulate between the retina and the choroid, andthe retina can become detached. In the wet (exudative) form, which ismore severe, blood vessels grow up from the choroid behind the retina,and the retina can also become detached. AMD can be treated with lasercoagulation, and with medication that stops and sometimes reverses thegrowth of blood vessels. AMD begins with characteristic yellow deposits(drusen) in the macula, between the retinal pigment epithelium and theunderlying choroid. Most patients with these early changes (referred toas age-related maculopathy) have good vision. Patients with drusen cango on to develop advanced AMD. The risk is considerably higher when thedrusen are large and numerous and associated with disturbance in thepigmented cell layer under the macula.

The term “macular edema” refers to the ocular diseases cystoid macularedema (CME) or diabetic macular edema (DME). CME is an ocular diseasethat affects the central retina or macula of the eye. When thiscondition is present, multiple cyst-like (cystoid) areas of fluid appearin the macula and cause retinal swelling or edema. CME may accompany avariety of diseases such as retinal vein occlusion, uveitis, and/ordiabetes. CME commonly occurs after cataract surgery. DME occurs whenblood vessels in the retina of patients with diabetes begin to leak intothe macula, the part of the eye responsible for detailed central vision.These leaks cause the macula to thicken and swell, progressivelydistorting acute vision. While the swelling may not lead to blindness,the effect can cause a severe loss in central vision.

The term “glaucoma” refers to an ocular disease in which the optic nerveis damaged in a characteristic pattern. This can permanently damagevision in the affected eye and lead to blindness if left untreated. Itis normally associated with increased fluid pressure in the eye (aqueoushumor). The term ocular hypertension is used for patients withconsistently raised intraocular pressure (IOP) without any associatedoptic nerve damage. Conversely, the term normal tension or low tensionglaucoma is used for those with optic nerve damage and associated visualfield loss but normal or low IOP. The nerve damage involves loss ofretinal ganglion cells in a characteristic patter. There are manydifferent subtypes of glaucoma, but they can all be considered to be atype of optic neuropathy. Raised intraocular pressure (e.g., above 21mmHg or 2.8 kPa) is the most important and only modifiable risk factorfor glaucoma, though some may have high eye pressure for years and neverdevelop damage, while others can develop nerve damage at a relativelylow pressure. Untreated glaucoma can lead to permanent damage of theoptic nerve and resultant visual field loss, which over time canprogress to blindness.

The term “uveitis” refers to an inflammatory disease of the uvea, thevascular layer of the eye sandwiched between the retina and the sclera(white of the eye). The uvea extends toward the front of the eye andconsists of the iris, choroid layer and ciliary body. Uveitis includesanterior uveitis, intermediate uveitis, and posterior uveitis. A commontype of uveitis is an inflammation of the iris called iritis (anterioruveitis). Uveitis may also occur at the posterior segment of the eye(e.g., at the choroid). Inflammation of the uvea can be recurring andcan cause serious problems such as blindness if left untreated, andaccounts for 10% of blindness globally. Early diagnosis and treatmentare important to prevent the complications of uveitis.

The term “dry eye” or “dry eyes” refers to an ocular disease in whichthere are insufficient tears to lubricate and nourish the eye. Tears arenecessary for maintaining the health of the front surface of the eye andfor providing clear vision. Patients with dry eyes either do not produceenough tears or have a poor quality of tears. Dry eye is a common andoften chronic problem, particularly in older adults. With each blink ofthe eyelids, tears are spread across the front surface of the eye, knownas the cornea. Tears provide lubrication, reduce the risk of eyeinfection, wash away foreign matter in the eye, and keep the surface ofthe eyes smooth and clear. When the normal amount of tear productiondecreases or tears evaporate too quickly from the eyes, symptoms of dryeye can develop. The most common form of dry eyes is due to aninadequate amount of the water layer of tears. This condition, calledkeratoconjunctivitis sicca (KCS), is also referred to as “dry eyesyndrome.” A non-limiting example of dry eye syndrome is Sjögren'ssyndrome.

The term “diabetic retinopathy” refers to retinopathy (i.e., a diseaseof the retina) caused by complications of diabetes, which can eventuallylead to blindness. Diabetic retinopathy may cause no symptoms, mildvision problems, or even blindness. Diabetic retinopathy is the resultof microvascular retinal changes. Hyperglycemia-induced intramuralpericyte death and thickening of the basement membrane lead toincompetence of the vascular walls. These damages change the formationof the blood-retinal barrier and also make the retinal blood vesselsbecome more permeable. Small blood vessels, such as those in the eye,are especially vulnerable to poor control over blood sugar. Anoveraccumulation of glucose and/or fructose damages the tiny bloodvessels in the retina. During the initial stage, called“nonproliferative diabetic retinopathy” (NPDR), most patients do notnotice any change in their vision. Early changes that are reversible anddo not threaten central vision are sometimes termed simplex retinopathyor background retinopathy. As the disease progresses, severenonproliferative diabetic retinopathy enters an advanced, “proliferativediabetic retinopathy” (PDR) stage when blood vessels proliferate. Thelack of oxygen in the retina causes fragile, new, blood vessels to growalong the retina and in the clear, gel-like vitreous humor that fillsthe inside of the eye, which may result in bleeding, cloudy vision,retina damage, or tractional retinal detachment.

The term “particle” refers to a small object, fragment, or piece of asubstance that may be a single element, inorganic material, organicmaterial, or mixture thereof. Examples of particles include polymericparticles, single-emulsion particles, double-emulsion particles,coacervates, liposomes, microparticles, nanoparticles, macroscopicparticles, pellets, crystals (e.g., crystalline forms of compounds oractive pharmaceutical agent), aggregates, composites, pulverized,milled, or otherwise disrupted matrices, and cross-linked protein orpolysaccharide particles. A particle may be composed of a singlesubstance or multiple substances. In certain embodiments, the particleis not a viral particle. In other embodiments, the particle is not aliposome. In certain embodiments, the particle is not a micelle. Incertain embodiments, the particle is substantially solid throughout. Incertain embodiments, the particle is a nanoparticle. In certainembodiments, the particle is a microparticle.

The term “nanoparticle” refers to a particle having a characteristicdimension in a nanometer range, such as less than about 1 micrometer andat least about 1 nanometer, where the characteristic dimension of theparticle is the smallest cross-sectional dimension of the particle. Acrystalline nanoparticle is referred to as a “nanocrystal.”

The terms “crystalline” or “substantially crystalline”, when used withrespect to nanostructures, refer to the fact that the nanostructurestypically exhibit long-range ordering across one or more dimensions ofthe structure. It will be understood by one of skill in the art that theterm “long range ordering” will depend on the absolute size of thespecific nanostructures, as ordering for a single crystal cannot extendbeyond the boundaries of the crystal. In this case, “long-rangeordering” will mean substantial order across at least the majority ofthe dimension of the nanostructure. In some instances, a nanostructurecan bear an oxide or other coating, or can be comprised of a core and atleast one shell. In such instances it will be appreciated that theoxide, shell(s), or other coating need not exhibit such ordering (e.g.it can be amorphous, polycrystalline, or otherwise). In such instances,the phrase “crystalline,” “substantially crystalline,” “substantiallymonocrystalline,” or “monocrystalline” refers to the central core of thenanostructure (excluding the coating layers or shells). The terms“crystalline” or “substantially crystalline” as used herein are intendedto also encompass structures comprising various defects, stackingfaults, atomic substitutions, and the like, as long as the structureexhibits substantial long range ordering (e.g., order over at leastabout 80% of the length of at least one axis of the nanostructure or itscore). In addition, it will be appreciated that the interface between acore and the outside of a nanostructure or between a core and anadjacent shell or between a shell and a second adjacent shell maycontain non-crystalline regions and may even be amorphous. This does notprevent the nanostructure from being crystalline or substantiallycrystalline as defined herein. The term “monocrystalline” when used withrespect to a nanostructure indicates that the nanostructure issubstantially crystalline and comprises substantially a single crystal.When used with respect to a nanostructure heterostructure comprising acore and one or more shells, “monocrystalline” indicates that the coreis substantially crystalline and comprises substantially a singlecrystal. When not used with respect to a nanostructure, the term“monocrystalline” to materials that are composed of substantially asingle crystallite of substantially the same size and orientation.

“Nanocrystal” is a nanostructure that is substantially monocrystalline.A nanocrystal thus has at least one region or characteristic dimensionwith a dimension in the nanometer range, such as less than about 1000nm, e.g., less than about 300 nm, less than about 200 nm, less thanabout 100 nm, or less than about 50 nm. Typically, the region orcharacteristic dimension will be along the smallest axis of thestructure. Optionally, a nanocrystal can comprise one or more surfaceligands (e.g., surfactants). The nanocrystal is optionally substantiallysingle crystal in structure (a “single crystal nanostructure” or a“monocrystalline nanostructure”). The term “nanocrystal” is intended toencompass substantially monocrystalline nanostructures comprisingvarious defects, stacking faults, atomic substitutions, and the like, aswell as substantially monocrystalline nanostructures without suchdefects, faults, or substitutions. In the case of nanocrystalheterostructures comprising a core and one or more shells, the core ofthe nanocrystal is typically substantially monocrystalline, but theshell(s) need not be.

The term “polycrystalline” refers to materials that are composed of manycrystallites of varying size and orientation. When used with respect tonanostructures, the term “polycrystalline” refers to a crystallinenanostructure that is not monocrystalline.

As used herein, the terms “pharmaceutical composition” and “formulation”are used interchangeably.

As used herein, the terms “pharmaceutical agent” and “drug” are usedinterchangeably.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

The present disclosure provides a compound having the formula

or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,and pharmaceutically acceptable salts and prodrugs thereof.

Unless otherwise indicated, reference to a compound, such as a compoundof Formula I, by structure or name includes alternate solid forms suchas solvates, hydrates, polymorphs, co-crystals; tautomers; andstereoisomers, such as diastereomers and enantiomers, includingdiastereomeric mixtures (both equal mixtures and mixtures enriched withone or more diastereomers) and enantiomeric mixtures (both racemicmixtures and mixtures having an enantiomeric excess).

In one embodiment, the compound of Formula (I) is formulated intomucus-penetrating particles or mucus-penetrating nanocrystals(collectively, MPPs) suitable for administration (e.g., topical orinhalation) to tissues of the subject having mucus (e.g., eye,respiratory tract, gastrointestinal tract, genito-urinary tract).

In another aspect, the present disclosure provides particles comprising1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea.In certain embodiments, the particles are nanoparticles. In one aspect,the nanoparticles are mucus penetrating. The particles may include acoating surrounding a core. The core may comprise primarily the compoundof Formula (I), or the core may be a polymeric core with the compoundencapsulated in the polymer. In certain embodiments, the particles arenanoparticles (e.g., particles having an average diameter of at leastabout 10 nm and less than about 1 μm). The particles may be useful indelivering the compound of Formula (I) to a subject. In certainembodiments, the particles are capable of delivering the compound in orthrough mucus of a subject.

In some embodiments, the compound of Formula (I) described herein may beintended for delivery in a subject's tissues having mucus (e.g., eye,respiratory tract, gastrointestinal tract, genito-urinary tract), whichis a viscoelastic and adhesive substance that traps most foreign objects(e.g., microorganisms, particles, dust). For effective drug delivery,compound or particles that are immobilized in the mucus are quicklyeliminated by mucus clearance mechanisms; therefore, they are not ableto effectively deliver the intended therapeutic effect. In thesetissues, for the compound to effective, it must quickly penetrate themucus and/or avoid mucus clearance mechanisms. Accordingly, modifyingmucoadhesive compounds or particles containing compounds with a coatingto reduce the mucoadhesiveness, and decreasing the size of the particlesof compound may allow for efficient delivery and therapeutic effect.

In one aspect, a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is formulated into mucus-penetrating particles or mucus-penetratingnanocrystals (collectively, MPPs) suitable for administration (e.g.,topical or inhalation) to tissues of the subject having mucus (e.g.,eye, respiratory tract, gastrointestinal tract, genito-urinary tract).

1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or mucus-penetrating particles or mucus-penetrating nanocrystalsthereof, may be suitable for being processed into mucus-penetratingpharmaceutical compositions (e.g., particles or crystals). In certainembodiments, the compound is suitable for milling (e.g., nano-milling).In certain embodiments, the compound is suitable for precipitation(e.g., microprecipitation, nanoprecipitation, crystallization, orcontrolled crystallization). In certain embodiments, the compound issuitable for emulsification. In certain embodiments, the compound issuitable for freeze-drying.

The present disclosure provides pharmaceutical compositions comprising acompound of Formula (I), or a pharmaceutically acceptable salt, solvate,hydrate, polymorph, co-crystal, tautomer, stereoisomer, or prodrugthereof, and a pharmaceutically acceptable excipient or carrier. Incertain embodiments, the pharmaceutical composition comprises a compoundof Formula (I), or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable excipient or carrier.

In another aspect, the present disclosure provides a pharmaceuticalcomposition comprising1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, and apharmaceutically acceptable excipient or carrier. In certainembodiments, the pharmaceutical composition comprises1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient or carrier.

In certain embodiments, the compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is provided in an effective amount in the pharmaceutical composition. Incertain embodiments, the effective amount is a therapeutically effectiveamount. In certain embodiments, the effective amount is aprophylactically effective amount. In certain embodiments, the effectiveamount is an amount effective for treating and/or preventing a disease.In certain embodiments, the effective amount is an amount effective fortreating a disease. In certain embodiments, the effective amount is anamount effective for treating and/or preventing a disease associatedwith aberrant signaling of a growth factor. In certain embodiments, theeffective amount is an amount effective for treating a diseaseassociated with aberrant signaling of a growth factor. In certainembodiments, the effective amount is an amount effective for treatingand/or preventing a disease associated with aberrant signaling ofvascular endothelial growth factor (VEGF). In certain embodiments, theeffective amount is an amount effective for treating a diseaseassociated with aberrant signaling of vascular endothelial growth factor(VEGF). In certain embodiments, the effective amount is an amounteffective for treating and/or preventing a disease associated withabnormal angiogenesis, such as cancer, benign neoplasm, atherosclerosis,hypertension, inflammatory disease, rheumatoid arthritis, maculardegeneration, an ocular neovascular disease, choroidalneovascularization, retinal neovascularization, neovascular glaucoma,and diabetic retinopathy. In certain embodiments, the effective amountis an amount effective to treat cancer (e.g., an ocular cancer). Incertain embodiments, the effective amount is an amount effective totreat macular degeneration.

An effective amount of a compound may vary from about 0.001 mg/kg toabout 1000 mg/kg in one or more close administrations for one or severaldays (depending on the mode of administration). In certain embodiments,the effective amount per dose varies from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about 750 mg/kg, from about 0.1mg/kg to about 500 mg/kg, from about 1.0 mg/kg to about 250 mg/kg, andfrom about 10.0 mg/kg to about 150 mg/kg.

An effective amount of a compound disclosed herein may inhibit abnormalangiogenesis and/or aberrant signaling of a growth factor by at leastabout 10%, at least about 20%, at least about 30%, at least about 40%,at least about 50%, at least about 60%, at least about 70%, at leastabout 80%, or at least about 90%. An effective amount of a compounddisclosed herein may inhibit abnormal angiogenesis and/or aberrantsignaling of a growth factor by less than about 90%, less than about80%, less than about 70%, less than about 60%, less than about 50%, lessthan about 40%, less than about 30%, less than about 20%, or less thanabout 10%. Combinations of the ranges described herein (e.g., at least20% and less than 50%) are also within the scope of the presentdisclosure. In certain embodiments, an effective amount of a compounddisclosed herein inhibits abnormal angiogenesis and/or aberrantsignaling of a growth factor by a percentage or a range of percentagedescribed herein, compared to normal angiogenesis and/or signaling.

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing the compound of Formula (I)described herein (i.e., the “active ingredient”) into association with acarrier or excipient, and/or one or more other accessory ingredients,and then, if necessary and/or desirable, shaping, and/or packaging theproduct into a desired single- or multi-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is a discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient, a compound of Formula (I) or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is generally equal to the dosage of the active ingredient which would beadministered to a subject and/or a convenient fraction of such a dosagesuch as, for example, one-half or one-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition will vary, depending upon the identity, size,and/or condition of the subject treated and further depending upon theroute by which the composition is to be administered. The compositionmay comprise between 0.1% and 100% (w/w) active ingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

A compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,and pharmaceutical compositions thereof provided herein, can beadministered by any route, including enteral (e.g., oral), parenteral,intravenous, intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by solutions, suspensions,drops, powders, ointments, gels, and/or creams), mucosal, nasal, buccal,sublingual; by intratracheal instillation, bronchial instillation,and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.Specifically contemplated routes are oral administration, intravenousadministration (e.g., systemic intravenous injection), regionaladministration via blood and/or lymph supply, topical administration,and/or direct administration to an affected site. In general, the mostappropriate route of administration will depend upon a variety offactors including the nature of the agent (e.g., its stability in theenvironment of the gastrointestinal tract), and/or the condition of thesubject (e.g., whether the subject is able to tolerate oraladministration). In certain embodiments, the compound or pharmaceuticalcomposition disclosed herein is suitable for topical administration tothe eye of a subject.

In one aspect, the pharmaceutical compositions are suitable for topicaladministration. In another aspect, the pharmaceutical compositions aresuitable for injection. In one embodiment, the pharmaceuticalcompositions comprising a compound of Formula (I) are suitable fordelivery to the eye. In another aspect, pharmaceutical compositionscomprising a compound of Formula (I) are suitable for oraladministration. In yet another aspect, the pharmaceutical compositionscomprising a compound of Formula (I) are suitable for inhalation.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension, or emulsion in a nontoxic parenterally acceptable diluent orsolvent. The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

Compositions for rectal or vaginal administration are typicallysuppositories that can be prepared by mixing an active ingredient asdescribed herein with suitable non-irritating excipients or carrierssuch as cocoa butter, polyethylene glycol, or a suppository wax that aresolid at ambient temperature but liquid at body temperature andtherefore melt in the rectum or vaginal cavity and release the activeingredient.

Solid dosage forms for oral administration include capsules, gelatincapsules, tablets, pills, powders, and granules. In such solid dosageforms, the active ingredient (e.g., a compound of Formula (I)) is mixedwith at least one inert, pharmaceutically acceptable excipient orcarrier known in the art, including without limitation fillers orextenders, binders, humectants, disintegrating agents, solutionretarding agents, absorption accelerators, wetting agents, absorbents,and lubricants, and mixtures thereof. In the case of capsules, tablets,and pills, the dosage form may include a buffering agent.

The active ingredient can be in a micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings, and othercoatings well known in the pharmaceutical formulating art.

Dosage forms for topical and/or transdermal administration of a compoundof Formula (I) may include solutions, suspensions, ointments, pastes,creams, lotions, gels, powders, sprays, inhalants, and/or patches.Generally, the active ingredient is admixed under sterile conditionswith a pharmaceutically acceptable carrier or excipient and/or anyneeded preservatives and/or buffers as can be required. Additionally,the present disclosure contemplates the use of transdermal patches,which often have the added advantage of providing controlled delivery ofan active ingredient to the body. Such dosage forms can be prepared, forexample, by dissolving and/or dispensing the active ingredient in theproper medium. Alternatively or additionally, the rate can be controlledby either providing a rate controlling membrane and/or by dispersing theactive ingredient in a polymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices such as thosedescribed in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288;4,270,537; 5,015,235; 5,141,496; and 5,417,662, all of which areincorporated by reference herein for all they teach regardingintradermal pharmaceutical delivery. Intradermal compositions can beadministered by devices which limit the effective penetration length ofa needle into the skin, such as those described in PCT publication WO99/34850 and functional equivalents thereof. Alternatively oradditionally, conventional syringes can be used in the classical mantouxmethod of intradermal administration. Jet injection devices whichdeliver liquid vaccines to the dermis via a liquid jet injector and/orvia a needle which pierces the stratum corneum and produces a jet whichreaches the dermis are suitable. Jet injection devices are described,for example, in U.S. Pat. Nos. 5,480,381; 5,599,302; 5,334,144;5,993,412; 5,649,912; 5,569,189; 5,704,911; 5,383,851; 5,893,397;5,466,220; 5,339,163; 5,312,335; 5,503,627; 5,064,413; 5,520,639;4,596,556; 4,790,824; 4,941,880; 4,940,460; and PCT publications WO97/37705 and WO 97/13537, all of which are incorporated by referenceherein for all they teach regarding jet injection devices. Ballisticpowder/particle delivery devices which use compressed gas to acceleratethe compound in powder form through the outer layers of the skin to thedermis are suitable.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi-liquid preparations such as solutions,suspensions, liniments, lotions, oil-in-water and/or water-in-oilemulsions such as creams, ointments, and/or pastes. Topicallyadministrable formulations may, for example, comprise from about 1% toabout 10% (w/w) of pharmaceutical agent (e.g., a compound of Formula(I)), although the concentration of the pharmaceutical agent can be ashigh as the solubility limit of the pharmaceutical agent in the solvent.Formulations for topical administration may further comprise one or moreof the additional ingredients described herein. In one embodiment,pharmaceutical compositions suitable for topical administration aresolutions or suspensions. In another embodiment, the solution orsuspension is in the form of eye drops.

A pharmaceutical composition can be prepared, packaged, and/or sold in aformulation suitable for pulmonary administration via the buccal cavity.Such a formulation may comprise dry particles that comprise the activeingredient and which have a diameter in the range from about 0.5 toabout 7 nanometers, or from about 1 to about 6 nanometers. Suchcompositions are conveniently in the form of dry powders foradministration using a device comprising a dry powder reservoir to whicha stream of propellant can be directed to disperse the powder and/orusing a self-propelling solvent/powder dispensing container such as adevice comprising the active ingredient dissolved and/or suspended in alow-boiling propellant in a sealed container. Such powders compriseparticles wherein at least 98% of the particles by weight have adiameter greater than 0.5 nanometers and at least 95% of the particlesby number have a diameter less than 7 nanometers. Alternatively, atleast 95% of the particles by weight have a diameter greater than 1nanometer and at least 90% of the particles by number have a diameterless than 6 nanometers. Dry powder compositions may include a solid finepowder diluent such as sugar and are conveniently provided in a unitdose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition disclosedherein. Another formulation suitable for intranasal administration is acoarse powder comprising the active ingredient and having an averageparticle from about 0.2 to 500 micrometers. Such a formulation isadministered by rapid inhalation through the nasal passage from acontainer of the powder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) to as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein.

A pharmaceutical composition disclosed herein can be prepared, packaged,and/or sold in a formulation for ophthalmic administration, such assolutions, suspensions, ointments, or gels. Such formulations may, forexample, be in the form of eye drops including, for example, a 0.1/1.0%(w/w) solution and/or suspension of the active ingredient (e.g., acompound of Formula (I)) in an aqueous or oily liquid carrier orexcipient. Such drops may further comprise buffering agents, salts,and/or one or more other of the additional ingredients described herein.Other ophthalmically-administrable formulations that are useful includethose which comprise the active ingredient in microcrystalline formand/or in a liposomal preparation. Eye drops are also contemplated asbeing within the scope of this disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions that aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

A compound of Formula (I) provided herein may be formulated in dosageunit form for ease of administration and uniformity of dosage. It willbe understood, however, that the total daily usage of the compositionsdisclosed herein will be decided by the attending physician within thescope of sound medical judgment. The specific therapeutically effectivedose level for any particular subject or organism will depend upon avariety of factors including the disease being treated and the severityof the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex, and diet of the subject; the time ofadministration, route of administration, and rate of excretion of thespecific active ingredient employed; the duration of the treatment;drugs used in combination or coincidental with the specific activeingredient employed; and like factors well known in the medical arts.

The exact amount of a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,required to achieve an effective amount will vary from subject tosubject, depending, for example, on species, age, and general conditionof a subject, severity of the side effects or disorder, identity of theparticular compound, mode of administration, and the like. The desireddosage can be delivered three times a day, two times a day, once a day,every other day, every third day, every week, every two weeks, everythree weeks, or every four weeks. In certain embodiments, the desireddosage can be delivered using multiple administrations (e.g., two,three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen, fourteen, or more administrations).

In certain embodiments, an effective amount of a compound of Formula (I)for administration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound of Formula (I) described herein maybe at dosage levels sufficient to deliver from about 0.001 mg/kg toabout 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, preferablyfrom about 0.1 mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kgto about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about0.1 mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg toabout 25 mg/kg, of subject body weight per day, one or more times a day,to obtain the desired therapeutic and/or prophylactic effect.

It will be appreciated that close ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

In certain embodiments, a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,pharmaceutical compositions thereof, and methods disclosed herein areuseful for applications in the eye, such as treating and/or preventingan ocular disease (e.g., macular degeneration, dry eye syndrome,diabetic macular edema, cystoid macular edema, uveitis, allergicconjunctivitis, glaucoma, and ocular rosacea). In certain embodiments,the pharmaceutical compositions disclosed herein can be topicallyadministered to the eye of a subject. Topical pharmaceuticalcompositions administered to the eye are advantageous overpharmaceutical compositions that are administered to the eye byinjection or orally.

In certain embodiments, a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is formulated as a mucus-penetrating particles. Methods for preparingmucus-penetrating particles have been described in, for example, U.S.Patent Publication Nos. 2008/0166414, 2010/0215580, 2013/0164343,2013/0236556, 2013/0316001, 2013/0316006, 2013/0316009, and2013/0323179, each of which is herein incorporated by reference for allthey teach regarding mucus penetrating particles.

In one embodiment, disclosed herein are pharmaceutical compositionscomprising a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,wherein said pharmaceutical composition comprises mucus-penetratingparticles of a compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,and a pharmaceutically acceptable excipient or carrier, and wherein saidmucus-penetrating particles have reduced inflammation uponadministration to a subject.

The compound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,is a potent inhibitor of VEGF receptor-2 (VEGFR2) and platelet-derivedgrowth factor receptor-β (PDGFR-β).

A compound of Formula (I) may be used in treating and/or preventing adisease (e.g., a disease associated with abnormal angiogenesis and/oraberrant signaling of a growth factor signaling pathway (e.g., VEGF)) ina subject in need thereof. The compounds disclosed herein may also beused in inhibiting abnormal angiogenesis and/or aberrant signaling of agrowth factor pathway in a subject and/or cell.

A range of diseases may result when the body of a subject loses controlover angiogenesis, i.e., new blood vessels grow abnormally (i.e.,excessively or insufficiently) or grow as a result of a tumor. Excessiveangiogenesis is often observed in subjects with diseases such asproliferative diseases (e.g., cancers, benign neoplasms, inflammatorydiseases, autoimmune diseases) and ocular diseases, especially withcancer, diabetic retinopathy, macular degeneration, rheumatoidarthritis, and psoriasis. In these diseases, new blood vessels feedabnormal tissues and/or destroy normal tissues. Excessive angiogenesismay occur when there are abnormal amounts of angiogenic growth factorspresent, overwhelming the effects of natural angiogenesis inhibitors.Therefore, inhibiting new blood vessel growth may be useful to treatdiseases associated with excessive angiogenesis. Insufficientangiogenesis is typically observed in subjects with a disease such ascoronary artery disease, stroke, or chronic wounds. In these diseases,blood vessel growth is inadequate, and circulation is not properlyrestored, which may lead to tissue death.

VEGFs have been found to play a major role in angiogenesis, for example,by increasing the number of capillaries in a given network. In vitrostudies have demonstrated that bovine capillary endothelial cellsproliferated and showed signs of tube structures upon stimulation withVEGF. Upregulation of VEGF is a major component of the physiologicalresponse to exercise and its role in angiogenesis is suspected to be apossible treatment in vascular injuries. In vitro studies have showedthat VEGFs are a potent stimulator of angiogenesis because, among otherthings, in the presence of this growth factor, plated endothelial cellswill proliferate and migrate, eventually forming tube structuresresembling capillaries. VEGFs may cause a massive signaling cascade inendothelial cells. Binding to VEGF2 starts a tyrosine kinase signalingcascade that stimulates the production of factors that variouslystimulate vessel permeability, proliferation/survival, migration, andfinally differentiation into mature blood vessels. Mechanically, VEGF isupregulated with muscle contractions as a result of increased blood flowto affected areas. The increased flow also causes a large increase inthe mRNA production of VEGF receptors 1 and 2. The increase in receptorproduction indicates that muscle contractions could cause upregulationof the signaling cascade relating to angiogenesis.

In one aspect, provided herein are methods of treating and/or preventinga disease associated with abnormal angiogenesis in a subject in needthereof. In certain embodiments, the disease being treated and/orprevented by the disclosed methods is associated with excessive and/orpathological angiogenesis.

In another aspect, provided herein are methods of treating and/orpreventing a disease associated with aberrant signaling of a growthfactor in a subject in need thereof. In certain embodiments, the diseaseis associated with excessive signaling of the growth factor. In certainembodiments, the disease being treated and/or prevented by the disclosedmethods is associated with aberrant signaling of VEGF. In certainembodiments, the disease is associated with excessive or aberrantsignaling of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-F, and/orplatelet-derived growth factor (PDGF).

In certain embodiments, the disease being treated and/or prevented bythe disclosed methods is a proliferative disease. All types ofproliferative diseases described herein may be treated and/or preventedby the disclosed methods. In certain embodiments, the disease beingtreated and/or prevented by the disclosed methods is cancer. All typesof cancer described herein may be treated and/or prevented by thedisclosed methods. In certain embodiments, the cancer is an ocularcancer. In certain embodiments, the ocular cancer is retinoblastoma,medulloepithelioma, uveal melanoma, ciliary body melanoma, or primaryintraocular lymphoma. In certain embodiments, the disease being treatedand/or prevented by the disclosed methods is a benign neoplasm. Alltypes of benign neoplasm described herein may be treated and/orprevented by the disclosed methods. In certain embodiments, the benignneoplasm is an ocular benign neoplasm. In certain embodiments, thebenign neoplasm is orbital dermoid cysts.

In certain embodiments, the disease being treated and/or prevented bythe disclosed methods is an inflammatory disease. All types ofinflammatory diseases described herein may be treated and/or preventedby the disclosed methods. In certain embodiments, the inflammatorydisease is an ocular inflammatory disease. In certain embodiments, theocular inflammatory disease is post-surgical inflammation. In certainembodiments, the disease being treated and/or prevented by the disclosedmethods is an autoimmune disease. All types of autoimmune diseasesdescribed herein may be treated and/or prevented by the disclosedmethods. In certain embodiments, the autoimmune disease is rheumatoidarthritis. In certain embodiments, the disease being treated and/orprevented by the disclosed methods is diabetes. In certain embodiments,the disease is type 1 diabetes. In certain embodiments, the disease istype 2 diabetes. In certain embodiments, the disease is gestationaldiabetes.

In some embodiments the disease being treated and/or prevented by thedisclosed methods is an ocular disease. In certain embodiments, theocular disease is an ocular neovascular disease. In some embodiments,the ocular disease being treated and/or prevented by the disclosedmethods is an anterior ocular disease that occurs at the anteriorportion or “front” of the eye of a subject. The anterior portion of theeye includes the cornea, iris, conjunctiva, tear film, cornealepithelium, anterior chamber, lens, ciliary body, ciliary zonule,posterior chamber, retina, macula, sclera, an optic nerve, choroid, andvitreous chamber. In certain embodiments, the anterior ocular diseasebeing treated and/or prevented by the disclosed methods is allergy,post-surgical inflammation, uveitis, an infection (e.g., a viral,bacterial, or fungal infection), aphakia, pseudophakia, astigmatism,blepharospasm, cataract, a conjunctival disease, conjunctivitis, acorneal disease, corneal oedema, meibomiam gland disease, cornealtransplant surgery, corneal ulcer, dry eye (e.g., dry eye syndrome), aneyelid disease, a lacrimal apparatus disease, lacrimal duct obstruction,laser induced exudation, myopia, presbyopia, pterygium, pupil disorders,corneal neovascularization, a refractive disorder, strabismus, orglaucoma.

In some embodiments, the ocular disease being treated and/or preventedby the disclosed methods is a posterior ocular disease that occurs atthe posterior portion or “back” of the eye. The posterior portion of theeye includes the choroid, sclera, vitreous humor, vitreous chamber,retina, macula, optic nerve, and blood vessels and nerves whichvascularize or innervate a posterior ocular region or site. In certainembodiments, the posterior ocular disease being treated and/or preventedby the disclosed methods is intraocular melanoma, acute macularneuroretinopathy, an exudative eye disease, Behcet's disease, exudativeretinopathy, macular oedema, retinopathy of prematurity, an epiretmalmembrane disorder, choroidal neovascularization, uveitis, diabeticuveitis, histoplasmosis, an infection (e.g., a viral, bacterial, orfungal infection), macular degeneration (e.g., acute maculardegeneration and age-related macular degeneration (AMD, such asnon-exudative age-related macular degeneration and exudative age-relatedmacular degeneration)), edema (e.g., macular edema, such as cystoidmacular edema (CME) and diabetic macular edema (DME)), multifocalchoroiditis, ocular trauma which affects a posterior ocular site orlocation, ocular cancer, a retinal disorder (e.g., central retinal veinocclusion), diabetic retinopathy (e.g., proliferative diabeticretinopathy and non-proliferative diabetic retinopathy), proliferativevitreoretinopathy (PVR), retinal arterial occlusive disease, retinaldetachment, uveitic retinal disease, sympathetic opthalmia, VogtKoyanagi-Harada (VKH) syndrome, uveal diffusion, a posterior ocularcondition caused by or influenced by an ocular laser treatment, aposterior ocular condition caused by or influenced by a photodynamictherapy, photocoagulation, radiation retinopathy, an epiretinal membranedisorder, branch retinal vein occlusion, anterior ischemic opticneuropathy, non-retinopathy diabetic retinal dysfunction, retinitispigmentosa, retinoblastoma, or glaucoma. In certain embodiments, theocular disease being prevented and/or treated by the disclosed methodsis macular degeneration. In certain embodiments, the ocular disease isage-related macular degeneration (AMD). In certain embodiments, theocular disease is glaucoma. In certain embodiments, the ocular diseaseis diabetic retinopathy. In certain embodiments, the ocular disease isretinoblastoma. In certain embodiments, the ocular disease is edema. Incertain embodiments, the ocular disease is cystoid macular edema (CME).In certain embodiments, the ocular disease is diabetic macular edema(DME). In certain embodiments, the ocular disease is an ocularinflammatory disease. In certain embodiments, the ocular disease ispost-surgical inflammation. In certain embodiments, the ocular diseaseis uveitis (e.g., anterior uveitis, intermediate uveitis, and postuveitis). In certain embodiments, the ocular disease is blepharitis. Incertain embodiments, the ocular disease is panuveitis. In certainembodiments, the ocular disease is scleritis. In certain embodiments,the ocular disease is dry eye. In certain embodiments, the oculardisease is Sjögren's syndrome. In certain embodiments, the oculardisease is an eye surgery.

Another aspect of the present disclosure relates to methods ofinhibiting the aberrant signaling of a growth factor signaling pathway(e.g., VEGF and/or PDGF) in a subject or cell.

In another aspect, the present disclosure provides methods of inhibitingabnormal or pathological angiogenesis in a subject in need thereof.

In one embodiment, the present disclosure relates to a method oftreating an ocular disease comprising administering to a subject in needthereof a therapeutically effective amount of a compound of Formula (I),or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, or apharmaceutical composition thereof.

In one aspect, the ocular disease is retinopathy. In another embodiment,the ocular disease is age-related macular degeneration (AMD). In anotherembodiment, the ocular disease is an ocular neovascular disease. Inanother embodiment, the ocular disease is corneal neovascularization. Inyet another embodiment, the ocular disease is diabetic macular edema(DME). In a further embodiment, the ocular disease is cystoid macularedema (CME). In yet another embodiment, the ocular disease is retinalvein occlusion (RVO).

In certain embodiments, the subject described herein is a human. Incertain embodiments, the subject is an animal. The animal may be ofeither sex and may be at any stage of development. In certainembodiments, the subject is a mammal. In certain embodiments, thesubject is a domesticated animal, such as a dog, cat, cow, pig, horse,sheep, or goat. In certain embodiments, the subject is a companionanimal such as a dog or cat. In certain embodiments, the subject is alivestock animal such as a cow, pig, horse, sheep, or goat. In certainembodiments, the subject is a zoo animal. In another embodiment, thesubject is a research animal such as a rodent (e.g., mouse, rat), dog,pig, or non-human primate. In certain embodiments, the animal is agenetically engineered animal. In certain embodiments, the animal is atransgenic animal.

In certain embodiments, the cell described herein is in vivo. In certainembodiments, the cell is in vitro. In certain embodiments, the cell isex vitro.

In certain embodiments, the methods of the present disclosure includeadministering to a subject in need thereof an effective amount ofcompound of Formula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, orpharmaceutical compositions thereof, as described herein. In certainembodiments, the methods include contacting a cell with an effectiveamount of a compound of Formula (I), or a pharmaceutical compositionthereof, as described herein.

In another aspect, the present disclosure relates to a compound ofFormula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, orpharmaceutical compositions thereof, for use in the treatment and/orprevention of a disease described herein in a subject in need thereof.

In yet another aspect, the present disclosure provides a compound ofFormula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, orpharmaceutical compositions thereof, for use in the inhibition ofabnormal angiogenesis in a subject in need thereof.

In still another aspect, the present disclosure provides compound ofFormula (I), or1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, or prodrug thereof, orpharmaceutical compositions thereof, for use in the inhibition ofaberrant signaling of a growth factor in a subject or cell in needthereof.

EXAMPLES

In order that the disclosure described herein may be more fullyunderstood, the following examples are set forth. The synthetic andbiological examples described in this application are offered toillustrate the compounds, pharmaceutical compositions, and methodsprovided herein and are not to be construed in any way as limiting theirscope.

Example 1 Preparation of Intermediate Compound 8

Compound 2

Lithium hexamethyldisilazane (436 mL, 0.44 mmol, 1 M solution inhexanes) was dissolved in diethyl ether (1 L). Compound 1,3-bromo-2-methylpyridine, (25.0 g, 0.14 mmol) was added. The solutionwas stirred for 1 h. Diethyl carbonate (26.0 mL, 0.22 mol) was added andthe solution was stirred overnight. The reaction solution was washedthree times with half-saturated aqueous sodium chloride (3×250 mL) anddried with magnesium sulfate. The solvent was evaporated and thereminder was dissolved in hexanes (200 mL). The solution was filteredthrough silica pad (10 g), the pad was rinsed with additional hexanes(100 mL) and the solvent was evaporated. The remaining oil was stirredin high vacuum for 1 hour till there were no further bubbles visible.The product was a yellow liquid (37.7 g). LCMS: [M+H]⁺=244.1.

Compound 3

Compound 2 (37.7 g, 0.15 mol) was dissolved in dry acetonitrile (400mL). 1,8-diazabicycloundec-7-ene (DBU) (28.2 g, 0.18 mol) was addedfollowed by 4-acetamidobenzenesulfonyl azide (37.6 g, 0.15 mol). Thesolution was stirred overnight. Water (3 L) was slowly added. The solidwas filtered off on a sintered glass funnel and washed with additionalwater (1 L) and hexanes (0.5 L) and air dried on the fritted funnel for2 h. The product was a cream solid (27.3 g). LCMS: [M+H]⁺=270.1.

Compound 4

Compound 3 (10.0 g, 37.0 mmol) and 4-nitrophenylboronic acid (10.0 g,59.9 mmol) were dissolved in dioxane (150 mL). Sodium carbonate (50 mL,2 M solution in water) and tetrakistriphenylphosphine (2.0 g, 1.73 mmol)were added. The suspension was degassed by passage of nitrogen (15 min).The solution was heated to 80° C. overnight. Water (500 mL) was addedand the precipitate was filtered off, washed with additional portion ofwater (500 mL) and hexanes (500 mL). The solid was air dried on asintered glass funnel for 2 hours and then dried in high vacuumovernight. The product was a yellow solid (9.6 g). LCMS: [M+H]⁺=313.1.

Compound 5

Compound 4 (9.6 g, 30.8 mmol) was dissolved in a mixture of dioxane (120mL), methanol (240 mL) and water (240 mL). Sodium hydroxide (42 mL, 10 Min water) was added and the solution was stirred for 6 h. The organicsolvents were evaporated. Water (600 mL) was added and the solution wasfiltered through a CELITE® pad. The filtrate was neutralized withhydrochloric acid (ca. 53 mL, 8 M in water) until the pH was 7. Theprecipitate was filtered off, washed with water (200 mL) and hexanes(200 mL). The solid was dried in high vacuum overnight. The product wasa cream solid (6.8 g). LCMS: [M+H]⁺=285.1.

Compound 6

Compound 5 (4.7 g, 16.5 mmol) was suspended in dichloromethane (100 mL).Oxalyl chloride (4.2 mL, 49.5 mmol) and N,N-dimethylformamide (50 μL)was added. The suspension was stirred for 1 h at which time all materialhas dissolved. The solvent was evaporated. The remainder wasco-evaporated with dichloromethane (2×100 mL) and dried in high vacuum(30 min). The red solid was dissolved in dry acetonitrile (150 mL).Sodium azide (9.4 g, 14 mmol) was added and the suspension was stirredfor 3 h. After this time the LCMS indicated full conversion. Water (1 L)was added and the cream solid was filtered off. The solid was dissolvedin trifluoroacetic acid (100 mL). Water (2 mL) was added and thesolution was heated at 60° C. for 2 h. After this time LCMS indicatedfull conversion. The solvent was evaporated and the reminder wasdissolved in acetonitrile (50 mL). The acetonitrile solution was slowlyadded to solution of sodium carbonate (500 mL, 2 M solution in water).The red precipitate was filtered off, washed with additional water (100mL) and hexanes (100 mL). The solid was air dried on a sintered glassfunnel for 2 hours and further dried in high vacuum for 3 h at 45° C.The product was a red solid (3.3 g). LCMS: [M+H]⁺=256.1. ¹H NMR(DMSO-d6): 8.78 (d, J=8.5 Hz, 1H), 8.34 (d, J=9.0 Hz, 2H), 8.07 (d,J=8.5 Hz, 1H), 7.72 (d, J=9.0 Hz, 2H), 7.08-7.02 (m, 1H), 4.57 (s, 2H).

Compound 7

Compound 6 (2.6 g, 10.1 mmol) was suspended in dichloromethane (40 mL)and acetonitrile (80 mL). Di-tert-butyl dicarbonate (5.6 g, 25.7 mmol)was added followed up by 4-dimethylaminopyridine (121 mg, 1.0 mmol). Thesuspension was stirred overnight at which point all the solids havedissolved. The brown solution was evaporated. The residue was coevaporated with dichloromethane (50 mL). The residue was dissolved indichloromethane (10 mL). Diethyl ether (300 mL) was added and thesolution was evaporated (to ca 50 mL). The solution was sonicated andhexanes (100 mL) were added. The precipitate was filtered off and driedin high vacuum for 2 h. The product was a yellow solid (4.0 g). LCMS:[M+Na]⁺=478.2.

Compound 8

Compound 7 (3.5 g, 7.69 mmol) was dissolved in methanol (250 mL). RaneyNickel (ca. 1 mL, Aldrich, RaNi 2800) was added followed by hydrazinehydrate (2.0 mL, 41.2 mmol). The solution was brought to a brief reflux.When the reflux stopped the LCMS indicated completion of the reaction.The catalyst was filtered off on a CELITE pad. The solvent wasevaporated to ca. 10 mL and the residue was diluted with water (200 mL).The solids were filtered off and the residue was washed with water (100mL) and hexanes (50 mL). The solid was dried in high vacuum overnight.The product was a cream solid (3.0 g). LCMS: [M+H]⁺=426.3. ¹H NMR(DMSO-d6): 8.61 (d, J=7.0 Hz, 1H), 7.28-7.25 (m, 2H), 7.13 (d, J=7.0 Hz,1H), 7.02 (dd, J=7.0, 7.0 Hz, 1H), 6.70 (d, J=9.0 Hz, 2H), 3.85 (s, 2H),1.31 (s, 18H).

Example 2 Synthesis of1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea

Compound 8,4-(4-Aminophenyl)-[1,2,3]triazolo[1,5-a]pyridin-3-yl-bis(tert-butylcarbamate) (200 mg, 0.48 mmol) was dissolved in dichloromethane (20 mL).Triethylamine (0.28 mL, 1.92 mmol) was added, the solution was cooled to−78° C. and phosgene (0.40 mL, 15% solution in toluene, 0.58 mmol) wasadded. The solution was stirred for 0.5 h and warmed up to roomtemperature. After 0.5 h 3-chloroaniline (185 mg, 1.46 mmol) was added.The solution was stirred for 2 h. The solvent was evaporated and theresidue was purified using flash chromatography (ISCO, 4 g column, 12column volumes, gradient from dichloromethane to ethyl acetate). Theproduct was a white solid (195 mg). LCMS: 7.54 min, 578.4 (M+, 40%),577.3 (M-1, 100%) (negative ionization mode).

The material from the previous step (195 mg, 0.34 mmol) was dissolved indichloromethane (0.5 mL). Trifluoroacetic acid (2 mL) was added and thesolution was stirred for 25 min. The solvent was evaporated (at roomtemperature). The residue was dissolved in dichloromethane (5 mL) andagain evaporated. The residue was dissolved in ethyl acetate (20 mL) andthe solution was washed with aqueous sodium bicarbonate. The solutionwas dried with magnesium sulfate and the solvent was evaporated toproduce Compound of Formula (I) (112 mg). LCMS: 6.42 min, 380.2 (M+2,10%), 379.2 (M+1, 30%), 157.1 (100%) (positive ionization mode). ¹H NMR(dmso-d6): 9.00 (d, J=3.1 Hz, 2H), 8.67 (d, J=7.1 Hz, 1H), 7.75-7.72 (m,1H), 7.62-7.67 (m, 2H), 7.44-7.51 (m, 2H), 7.29-2.37 (m, 2H), 7.01-7.09(m, 1H), 6.97 (dd, J=6.4, 24.6 Hz, 2H), 4.45 (s, 2H).

Example 3 Crystalline Form of Compound I Preparation of Crystalline FormI

Crystal form I was prepared by stirring a slurry of compound I in ethylacetate.

In one instance, 30.56 mg of compound I was deposited into a 4 mLscintillation vial with screw-top, henceforth referred to as the workingvial. To the working vial was added a 12×4.5 mm magnetic stir bar alongwith 1 mL room temperature ethyl acetate. The vial was capped andtransferred into an IKA stirring plate equipped with a magnetic stirrerthat was set at room temperature. The suspension in the vial was allowedto stir at room temperature and 500 rpm for 5 days.

After 5 days stirring at room temperature, the solid material wascollected by centrifuge filtration and the filtrate was discarded. Thecentrifuge filter tube was lightly covered then dried under high vacuumfor approximately 17 hours. The recovered dried solid was analyzed byXRPD, which showed a unique powder pattern that was assigned as crystalform I.

In another instance, 30.55 mg of compound I was deposited into a 4 mLscintillation vial with screw-top, henceforth referred to as the workingvial. To the working vial was added a 12×4.5 mm magnetic stir bar alongwith 1 mL room temperature ethyl acetate. The vial was capped andtransferred into a hot plate equipped with a magnetic stirrer that waspreheated to 60° C. The suspension in the vial was allowed to stir at60° C. and 500 rpm for 5 days.

After 5 days stirring on hot plate, the solid material was collected bycentrifuge filtration and the filtrate was discarded. The centrifugefilter tube was lightly covered then dried under high vacuum forapproximately 17 hours. The recovered dried solid was analyzed by XRPD,which showed the powder pattern of crystal form I.

Properties of Crystalline Form I

For XRPD, patterns were obtained using a Rigaku MiniFlex 600 benchtopx-ray diffractometer equipped with a Cu X-ray tube (Cu/Kα=1.54059 {acuteover (Å)}), a six-position sample changer and a D/teX Ultra detector.

Sample Preparation of Neat Crystalline Form for XRPD, Procedure A

As described herein for the characterization of neat crystal Form I,milligram amounts of solid sample were firmly packed in the 5-mm×0.2-mmdepression of a zero background sample holder (Rigaku 906166 5 mm×0.2 mmWell, Si510).

Sample Preparation of Milled Crystals for XRPD, Procedure B

As described herein for the characterization of milled crystal form I,particles were isolated from bulk formulation by centrifugation at55,000 rpm over 15 minutes and deposited thinly and evenly onto a flatzero background XRPD sample holder (Rigaku 906165 Flush, Si510). Thesample was allowed to dry under gentle air stream, usually for up to 3minutes, until it was visually dry.

XRPD patterns were acquired from 3-40° two theta at 0.02° step size and5°/min scan speed using the following instrument settings: 40 kV-15 mAX-ray generator, 2.5° Soller Slit, 10 mm HIS, 0.625° Divergence Slit, 8mm Scatter Slit with Kβ filter, and an open Receiving Slit. Diffractionpatterns were viewed and analyzed using PDXL analysis software providedby the instrument manufacturer. Using Procedure A, a reference standardsilicon powder (NIST Standard Reference Material 640d) generated a peakat 28.39° two theta.

For DSC, about 2 mg of sample was weighed into a standard aluminumsample pan. The sample pan was loaded into the apparatus (Q1000Differential Scanning Calorimeter, TA Instruments), which was equippedwith an auto-sampler. A thermogram was obtained by individually heatingthe sample at a rate of 10° C./min from room temperature toapproximately 250-300° C. using an empty standard aluminum pan as areference. Dry nitrogen was used as a sample purge gas and was set toflow at 50 mL/min. Thermal transitions were viewed and analyzed usingthe analysis software provided with the instrument.

For TGA, about 3 mg of the sample was transferred into an aluminumsample pan. The pan was placed in the loading platform and was thenautomatically loaded into the apparatus (Q500 ThermogravimetricAnalyzer, TA Instruments) using the control software. Thermograms wereobtained by individually heating the sample at 10° C./min from roomtemperature to 300° C. under flowing dry nitrogen, with a sample purgeflow rate of 25 mL/min and a balance purge flow rate of 10 mL/min.Thermal transitions (e.g., weight changes) were viewed and analyzedusing the analysis software provided with the instrument.

The XRPD pattern of crystal form I is illustrated in FIG. 1. Atabulation of the peaks in two-theta and the corresponding d-spacingvalues comprised in the XRPD patter of form I are listed in Table 1.

TABLE 1 XRPD peak listing of neat crystal form I Position ± 0.2°d-spacing ± 0.2 Relative Intensity Peak No. [2θ] [Å] [%] 1 8.05 10.984.96 2 10.54 8.39 15.44 3 11.53 7.67 1.86 4 12.24 7.22 4.43 5 14.36 6.1635.85 6 14.83 5.97 9.77 7 15.82 5.60 12.44 8 16.16 5.48 20.38 9 16.485.37 3.20 10 17.16 5.16 4.98 11 19.26 4.60 17.00 12 20.20 4.39 23.68 1320.56 4.32 5.67 14 21.28 4.17 6.86 15 21.67 4.10 40.30 16 22.46 3.952.720 17 23.13 3.84 3.32 18 24.06 3.70 10.18 19 24.72 3.60 100.00 2025.27 3.52 26.26 21 25.99 3.43 26.22 22 26.23 3.40 52.39 23 26.77 3.3313.18 24 27.64 3.22 4.30 25 28.96 3.08 19.91 26 29.44 3.03 12.11 2730.36 2.94 12.86 28 31.23 2.86 8.09 29 32.43 2.76 8.69 30 34.02 2.633.27 31 34.92 2.57 6.67 32 35.88 2.50 5.59

FIG. 2 shows that the DSC thermogram measured from 25° C. to 300° C.,ramped at 10° C./min, was found to decompose without melting beyond 170°C. FIG. 3 shows that the TGA thermogram measured from 25° C. to 300° C.,ramped at 10° C./min, was also found to decomposes above 170° C. The DSCand TGA thermograms both indicate that crystal form I is an anhydrateform that decomposes above 170° C.

Example 4 Mucus Penetrating Particles (MPP) of Crystal Form I ofCompound I

A wet milling process was employed to generate MPP. Milling media,specifically 0.5 mL bulk volume of 1-mm ceria-stabilized zirconium oxidebeads, was added to a glass vial. Approximately 25 mg of Compound I and0.475 mL of milling solution, 5% (final w/w) PLURONIC® F127 in PBS(0.0067 M (PO₄ ³⁻) phosphate buffered saline pH 7.2), were then added tothe vial yielding a slurry of 5% Compound I (w/v). A magnetic stir barwas used to agitate the beads, stirring at approximately 500 rpm.Particle size after milling is listed in Table 2.

TABLE 2 Examples of formulating MPP of Compound I crystal form I.Milling Time Particle Size Starting Form (days) (nm) MPP Form crystalform I 3 190 crystal form I

After milling, XRPD patterns of Compound I with excipients still presentwere collected using Procedure B. Neat crystal form I is stable whenformulated as an MPP; details are listed in Table 2. FIG. 4 shows XRPDpatterns, which demonstrate that the crystalline form was unchangedafter milling.

Example 5 Cellular Assays: Vascular Endothelial Growth Factor Receptor 2(VEGFR2) and Platelet-Derived Growth Factor Receptor-β (PDGFR-β)

The human umbilical endothelial (HUE) cell line expresses high levels ofVEGFR2. The compound prepared in Example 2 above,1-(4-(3-amino-[1,2,3]triazolo[1,5-a]pyridin-4-yl)phenyl)-3-(3-chlorophenyl)urea,was tested for the ability to inhibit tyrosine auto-phosphorylation ofthis receptor after stimulation with vascular endothelial growth factorA (VEGF-A). HUE cells were plated into multi-well cell culture plates inendothelial cell growth medium (ECGM) supplemented with 10% fetal calfserum (FCS). After starvation in endothelial cell basal medium (ECBM)supplemented with 10% FCS overnight cells were incubated with compoundsin serum-free ECBM. On the day of the assay the medium was changed toserum-free ECBM, the compound was 1:3 serial diluted from 10 mM stock in100% DMSO and then 1:100 diluted to each well of cells in ECBM withoutFCS to create final solutions containing 1% DMSO and an eight pointcompound close response curve starting at 10⁻⁷ M, 10⁻⁸ M, or 10⁻⁹ M. Thecompound was added in duplicate to the multi-well cell culture plate.For controls, cells in one column of wells were treated with 1% DMSO toserve as High Control, and a second column of wells was treated with10⁻⁵ M staurosporine to serve as Low Control for the assay. Cells wereincubated for 90 min at 37° C. in ECBM with the serial diluted compoundand then stimulated for 3 min with 100 ng/ml VEGF-A. Cells were lysed,and then levels of phosphorylated VEGFR2 were determined in a sandwichELISA assay formatted in a multi-well plate using a VEGFR2 specificcapture antibody and an anti-phosphotyrosine VEGFR2 detection antibody.The raw data were converted to percent tyrosine phosphorylated VEGFR2with respect to the on plate High and Low Controls. IC₅₀ values weredetermined by fitting the converted eight point close response data to afour-parameter logistic equation using GRAPHPAD PRISM® 5.01 software.

The murine embryonal fibroblast cell line NIH3T3 expresses endogenouslyhigh levels of PDGFR-β. The compound prepared in Example 2 above wastested for the ability to inhibit tyrosine auto-phosphorylation of thisreceptor after stimulation with platelet-derived growth factor BB(PDGF-BB). The NIH3T3 cells were plated into multi-well cell cultureplates in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10%FCS. The medium was removed and then replaced with DMEM without FCS andcells serum starved overnight at 37° C. On the day of the assay thecompound was 1:3 serial diluted from 10 mM stocks in 100% DMSO and thenwere 1:100 diluted to each well of cells in DMEM without FCS to createfinal solutions containing 1% DMSO and an eight point compound closeresponse curve starting at 10⁻⁶ M, 10⁻⁷ M, or 10⁻⁸ M. The compound wasadded in duplicate to columns of the multi-well plate. Cells in onecolumn of wells of the multi-well plate were treated with 1% DMSO toserve as High Control, and cells in a second column of wells weretreated with 10⁻⁵ M staurosporine to serve as Low Control for the assay.Cells were incubated for 90 min at 37° C. in DMEM with the serialdiluted compound and then stimulated for 3 min with 100 ng/ml PDGF-BB.Cells were lysed, and then levels of phosphorylated PDGFR-β weredetermined in a sandwich ELISA assay formatted in a multi-well plateusing a PDGFR-β specific capture antibody and an anti-phosphotyrosinePDGFR-β detection antibody. The raw data were converted to percentphosphorylated PDGFR-β with respect to the on plate High and LowControls. IC₅₀ values were determined by fitting the converted eightpoint close response data to a four-parameter logistic equation usingGRAPHPAD PRISM® 5.01 software.

The results of these cellular assays indicated that the compound ofFormula (I) is a potent inhibitor of VEGFR2 and PDGFR-1 with cellularIC₅₀ values of 0.37 nM and 3.2 nM, respectively.

Example 6 Pharmacokinetic (PK) Study in Pigs

A pharmacokinetic (PK) study of a compound of Formula (I) formulated asa mucus-penetrating particles or nanocrystals (MPP) is performed inorder to demonstrate topical instillation results in drug exposure atthe back of the eye.

For example, pigs (such as Gottingen mini-pigs or Sinclair pigs) areused to perform a PK study. Animals receive a single topical ocularclose (e.g., a close volume of 35 μL) in the right eye twice daily(BID), approximately 12 hr apart (±1 hour), for 4 consecutive days; onthe fifth day animals will receive a single topical ocular close in themorning only for a total of 9 closes over the study duration.

All animals are euthanized with sodium pentobarbital and blood iscollected via cardiac puncture into tubes containing K₂EDTA, which iscentrifuged to obtain plasma. Then, both eyes are enucleated, flashfrozen and stored at −70° C. for at least 2 hr. Within approximately 2days, the frozen matrices are collected as right and left eye forchoroid and retina. Drug exposures in plasma and back of the eye arethen determined.

EQUIVALENTS AND SCOPE

In the claims articles such as “a,” “an,” and “the” may mean one or morethan one unless indicated to the contrary or otherwise evident from thecontext. Claims or descriptions that include “or” between one or moremembers of a group are considered satisfied if one, more than one, orall of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The invention includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Theinvention includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process.

Furthermore, the invention encompasses all variations, combinations, andpermutations in which one or more limitations, elements, clauses, anddescriptive terms from one or more of the listed claims is introducedinto another claim. For example, any claim that is dependent on anotherclaim can be modified to include one or more limitations found in anyother claim that is dependent on the same base claim. Where elements arepresented as lists, e.g., in Markush group format, each subgroup of theelements is also disclosed, and any element(s) can be removed from thegroup. It should it be understood that, in general, where the invention,or aspects of the invention, is/are referred to as comprising particularelements and/or features, certain embodiments of the invention oraspects of the invention consist, or consist essentially of, suchelements and/or features. For purposes of simplicity, those embodimentshave not been specifically set forth in haec verba herein. It is alsonoted that the terms “comprising” and “containing” are intended to beopen and permits the inclusion of additional elements or steps. Whereranges are given, endpoints are included. Furthermore, unless otherwiseindicated or otherwise evident from the context and understanding of oneof ordinary skill in the art, values that are expressed as ranges canassume any specific value or sub-range within the stated ranges indifferent embodiments of the invention, to the tenth of the unit of thelower limit of the range, unless the context clearly dictates otherwise.

This application refers to various issued patents, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference. If there is a conflict between any ofthe incorporated references and the instant specification, thespecification shall control. In addition, any particular embodiment ofthe present invention that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Because such embodimentsare deemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the invention can be excluded from any claim,for any reason, whether or not related to the existence of prior art.

Those skilled in the art will recognize or be able to ascertain using nomore than routine experimentation many equivalents to the specificembodiments described herein. The scope of the present embodimentsdescribed herein is not intended to be limited to the above Description,but rather is as set forth in the appended claims. Those of ordinaryskill in the art will appreciate that various changes and modificationsto this description may be made without departing from the spirit orscope of the present invention, as defined in the following claims.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt or prodrug thereof.
 2. A crystalform of

wherein the crystal form has an X-ray powder diffraction (XRPD) patternwith peaks at about 10.5, about 14.4, about 19.3, about 21.7, and about24.7 degrees
 29. 3. A pharmaceutical composition comprising a compoundof claim 1, or a pharmaceutically acceptable salt or prodrug thereof, ora crystal form of claim 2, and a pharmaceutically acceptable excipientor carrier.
 4. The pharmaceutical composition of claim 3, wherein thepharmaceutical composition is suitable for topical administration,injection, oral administration, or inhalation.
 5. (canceled)
 6. Thepharmaceutical composition of claim 3, wherein the pharmaceuticalcomposition is suitable for delivery to the eye.
 7. The pharmaceuticalcomposition of claim 6, wherein the pharmaceutical composition is in acontainer configured to administer eye drops.
 8. (canceled) 9.(canceled)
 10. A method of treating a disease comprising administeringto a subject in need thereof a therapeutically effective amount of apharmaceutical composition of claim
 3. 11. The method of claim 10,wherein the disease is a proliferative disease, or an ocular disease.12. The method of claim 11, wherein the disease is cancer. 13.(canceled)
 14. The method of claim 13, wherein the ocular disease isretinopathy.
 15. The method of claim 13, wherein the ocular disease isage-related macular degeneration (AMD).
 16. The method of claim 13,wherein the ocular disease is corneal neovascularization.
 17. The methodof claim 13, wherein the ocular disease is an ocular neovasculardisease.
 18. The method of claim 13, wherein the ocular disease isdiabetic macular edema.
 19. The method of claim 13, wherein the oculardisease is cystoid macular edema.
 20. The method of claim 13, whereinthe ocular disease is retinal vein occlusion.
 21. A method of inhibitinggrowth factor signaling comprising administering to a subject atherapeutically effective amount of a pharmaceutical composition ofclaim
 3. 22. The method according to claim 21, wherein the compound orthe pharmaceutical composition is administered topically, by injection,orally, or by inhalation.
 23. (canceled)
 24. The method according toclaim 21, wherein the compound or the pharmaceutical composition isadministered to the eye.
 25. (canceled)
 26. (canceled)
 27. (canceled)